Estrogenic Endocrine Disrupting Compounds Research Articles

Female-biased sex ratios and delayed puberty in two fish species with different Ecologies in an Anthropogenically affected urban lake

In aquatic ecosystems, endocrine-disrupting compounds (EDCs) pose a growing concern for their potential adverse effects on fish reproduction and development. In lake Pyhäjärvi, located in the urban boreal region of Tampere, Finland, a significant number of sexually immature pikeperch (Sander lucioperca) individuals have been identified in size and age categories that are expected to be sexually mature. To explore if this phenomenon is attributed to estrogenic endocrine disruption, we conducted a comprehensive study comparing fish from lake Pyhäjärvi with those from a nearby reference lake, lake Näsijärvi. Roach (Rutilus rutilus), known for its susceptibility to EDCs, was also included for comparison. We examined various parameters in both pikeperch and roach, including size, condition factor, age, reproductive indicators, biometric indices and gonadal histology. We also assessed liver vitellogenin mRNA levels and genetic sex in roach, and measured estrogen levels in lake waters and wastewater treatment plant effluents. Results revealed that approximately one-third of fish in both species exhibited sexual immaturity in lake Pyhäjärvi, with a female-biased sex ratio. Surprisingly, we found no signs of estrogenic endocrine disruption, indicated by the absence of intersex fish in both species. Furthermore, vitellogenin levels in roach closely resembled those in the reference lake. Estrogens were undetectable in the lake waters, suggesting that factors other than estrogenic EDCs, including other potential endocrine disruptors such as PCBs or heavy metals, may be influencing delayed sexual maturity and skewed sex ratios. Further inquiry is needed to pinpoint these underlying causes. Our study provides essential baseline information on fish sexual development in lake Pyhäjärvi, emphasizing the need for ongoing monitoring and research to understand delayed sexual maturity and biased sex ratios. This is vital given the increasing concern about EDC impacts on aquatic ecosystems and the necessity for effective management strategies to protect these ecosystems' health and integrity.

Endocrine-disrupting compounds and metabolomic reprogramming in breast cancer.

Endocrine-disrupting chemicals pose a growing threat to human health through their increasing presence in the environment and their potential interactions with the mammalian endocrine systems. Due to their structural similarity to hormones like estrogen, these chemicals can interfere with endocrine signaling, leading to many deleterious effects. Exposure to estrogenic endocrine-disrupting compounds (EDC) is a suggested risk factor for the development of breast cancer, one of the most frequently diagnosed cancers in women. However, the mechanisms through which EDCs contribute to breast cancer development remain elusive. To rapidly proliferate, cancer cells undertake distinct metabolic programs to utilize existing nutrients in the tumor microenvironment and synthesize macromolecules de novo. EDCs are known to dysregulate cell signaling pathways related to cellular metabolism, which may be an important mechanism through which they exert their cancer-promoting effects. These altered pathways can be studied via metabolomic analysis, a new advancement in -omics technologies that can interrogate molecular pathways that favor cancer development and progression. This review will summarize recent discoveries regarding EDCs and the metabolic reprogramming that they may induce to facilitate the development of breast cancer.

Polyaniline Entrapped Water-Dispersible 3MPA-ZnSe Quantum Dots and Their Application for the Development of an Enzymatic Electrochemical Nanobiosensor for the Detection of 17β-Estradiol, an Endocrine-Disrupting Compound.

17β-estradiol is used as a growth and fertility stimulant in the agronomic sector to induce fertility and manipulate reproductive characteristics in animals. However, unintended or unregulated distribution and exposure to even significant low levels of 17β-estradiol estrogen have detrimental health implication that can lead to reproductive abnormalities and even cancer. This could have severe effect on the ecosystem imbalance, food safety, to such a degree that its health impact necessitates rapid methods to probe for its prevalence and occurrence in the environment. Herein a simple, robust, sensitive and once-off use electrochemical biosensor to detect 17β-estradiol is developed, using 3-mercaptopropionic acid capped zinc selenide quantum dots trapped within the polyaniline (PANI) framework structure. The biosensor's interaction with the substrate was based on the capability of the hemeprotein, horseradish peroxidase (HRP) enzyme (i.e., baroreceptor) to alternatively catalyze phenolic alcohols. The biosensor displayed a significantly low limit of detection limit (LOD) of value 0.2 × 10-6M towards 17β-estradiol. The Mechaelis-Menten constant (Km) with the magnitude of 0.64 × 10-6M was obtained; this indicates an outstanding affinity of the biosensing films towards 17β-estradiol. Subsequently, the developed biosensor was able to accurately and efficiently measure successive concentrations of 17β-estradiol from 0.2 × 10 to 4 × 10-6M. The fabricated biosensor showed good selectivity towards 17β-estradiol compared to the other estrogenic endocrine-disrupting compounds such as estrone (E1), ethnylstradiol (EE2), and estriol (E3). The biosensor was capable of detecting 17β-estradiol in spiked tap water samples with good recoveries, thus affirming its potential to be applied for real electro-analysis of 17β-estradiol in treated wastewater.

Declines in Reproductive Condition of Male Largemouth Bass (Micropterus salmoides) Following Seasonal Exposure to Estrogenic Endocrine-Disrupting Compounds.

Reproductive abnormalities, that could lead to possible effects at the population level, have been observed in wild fish throughout the United States, with high prevalence in largemouth bass (LMB; Micropterus salmoides) and smallmouth bass (Micropterus dolomieu). Estrone (E1) and atrazine (ATR) are common environmental contaminants often associated with agricultural land use. 17alpha-ethinylestradiol (EE2) is a contaminant associated with wastewater treatment effluent, and a representative, well-studied estrogen commonly used for fish toxicity testing. Our objective was to assess whether early gonad recrudescence in adult fish was a period of sensitivity for alterations in reproductive condition and function. Adult male LMB were exposed from post-spawning to early gonad recrudescence to either a mixture of E1 (47.9 ng/L) + ATR (5.4 µg/L), or EE2 (2.4 ng/L) in outdoor experimental ponds. Gonad samples were collected from fish just prior to the start of exposure (July), at the end of the exposure period (December), the following spring just prior to spawning (April), and post spawning (May). Gonadosomatic index (GSI) was significantly reduced in E1 + ATR-exposed and EE2-exposed males compared to control at every post-exposure time point. Reduced sperm count and sperm motility were observed in the mixture treatment (E1 + ATR) compared to the control. Sperm motility was also reduced in the EE2 treatment. These data together indicate that estrogenic endocrine-disrupting compounds can lessen the reproductive condition of adult male LMB, and that effects of exposure during early gonad recrudescence can persist at least through the subsequent spawning cycle.

Exposure to 17α-Ethinylestradiol Results in Differential Susceptibility of Largemouth Bass (Micropterus salmoides) to Bacterial Infection.

Disease outbreaks, skin lesions, mortality events, and reproductive abnormalities have been observed in wild populations of centrarchids. The presence of estrogenic endocrine disrupting compounds (EEDCs) has been implicated as a potential causal factor for these effects. The effects of prior EEDC exposure on immune response were examined in juvenile largemouth bass (Micropterus salmoides) exposed to a potent synthetic estrogen (17α-ethinylestradiol, EE2) at a low (EE2Low, 0.87 ng/L) or high (EE2High, 9.08 ng/L) dose for 4 weeks, followed by transfer to clean water and injection with an LD40 dose of the Gram-negative bacteria Edwardsiella piscicida. Unexpectedly, this prior exposure to EE2High significantly increased survivorship at 10 d post-infection compared to solvent control or EE2Low-exposed, infected fish. Both prior exposure and infection with E. piscicida led to significantly reduced hepatic glycogen levels, indicating a stress response resulting in depletion of energy stores. Additionally, pathway analysis for liver and spleen indicated differentially expressed genes associated with immunometabolic processes in the mock-injected EE2High treatment that could underlie the observed protective effect and metabolic shift in EE2High-infected fish. Our results demonstrate that exposure to a model EEDC alters metabolism and immune function in a fish species that is ecologically and economically important in North America.

Abstract 2687: The estrogenic activities of endocrine disruptors alter the extracellular matrix and tissue stiffness in the mammary gland

Abstract In utero exposure to estrogenic endocrine disrupting compounds (EDCs) increases a woman’s lifetime risk of breast cancer. Similarly, mice exposed in utero to the estrogenic EDC bisphenol A (BPA) have increased susceptibility to mammary gland tumors. It is unclear which BPA-induced alterations predispose the mammary gland to cancer transformation. There is a critical need to understand the mechanisms that drive increased cancer risk in order to assess the impact of BPA and BPA alternatives that retain estrogenic activity. We have utilized in utero BPA exposure as a model system for in utero estrogenic endocrine disruption to study the long-term consequences to the mouse mammary stroma. We found that BPA exposed fibroblasts showed significant transcriptional deregulation, with the extracellular matrix being the most altered cellular component and multiple collagen genes being more highly expressed. The fibroblasts from the BPA exposed mice decreased fluid permeability of the extracellular matrix, indicative of an increased density in the extracellular matrix. Also, in utero BPA exposure increased mammary gland stiffness. Changes to breast density, stiffness, and collagen deposition are all associated with breast cancer risk. Further, we test BPA alternative compounds with varying affinities for the estrogen receptor in our in utero model to assess the phenotypes in the mouse mammary stroma which are associated with breast cancer risk. Additionally, we use a mesenchymal estrogen receptor alpha (ERα) knockout mouse model to dissect the in utero cellular target of EDCs. Citation Format: Clarissa Wormsbaecher, Andrea R. Hindman, Alex Avendano, Marcos Cortes-Medina, Jonathan W. Song, Craig J. Burd. The estrogenic activities of endocrine disruptors alter the extracellular matrix and tissue stiffness in the mammary gland [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2687.

Urinary Concentrations of Triclosan, Bisphenol A, and Brominated Flame Retardants and the Association of Triclosan with Demographic Characteristics and Body Fatness among Women with Newly Diagnosed Breast Cancer.

Background: Triclosan, bisphenol A (BPA), and brominated flame retardants are environmental estrogenic endocrine-disrupting compounds that may influence the prognosis of breast cancer. We examined the urinary concentrations of these compounds and their associations with demographic characteristics and body fatness in a population of women with newly diagnosed breast cancer. Methods: Overnight urine collection and anthropometric measures were obtained from 302 participants. Triclosan, BPA, tetrabromobisphenol A (TBBPA), and tetrabromobenzoic acid (TBBA) concentrations were determined using ultra-performance liquid chromatography–tandem mass spectrometry. Regression analyses were conducted to examine associations of urinary compound concentration with age, menopause, race, ethnicity, educational level, estrogen receptor status, body size, and body composition. Results: Triclosan, BPA, and TBBA were detected in urine samples from 98.3%, 6.0%, and 0.3% of patients, respectively; TBBPA was undetectable. Among patients with quantifiable values, the geometric mean concentrations were 20.74 µg/L (27.04 µg/g creatinine) for triclosan and 0.82 µg/L (1.08 µg/g creatinine) for BPA. Body mass index ≥ 30 vs. <25 kg/m2 was associated with lower creatinine-corrected urinary concentrations of triclosan (−40.00, 95% confidence interval [CI] = −77.19 to −2.81; p = 0.0351). The observed association was predominantly in postmenopausal women (−66.57; 95% CI: −109.18% to −23.96%). Consistent results were found for associations between triclosan levels and fat mass variables. Conclusion: In this study population, women with newly diagnosed breast cancer had triclosan exposure. Assessments of the implications of urinary concentrations of triclosan for women should consider body fatness and menopausal status.

Estrogens as immunotoxicants: 17α-ethinylestradiol exposure retards thymus development in zebrafish (Danio rerio).

Estrogenic endocrine disrupting compounds (EEDCs) can cause alterations in sexual development and reproductive function of fish. Growing evidence suggests that EEDCs can also interfere with development and function of innate immunity of fish. The present study examined a potential disruptive effect of EEDCs at field-relevant concentrations on the development of adaptive immunity, more specifically the thymus. Zebrafish (Danio rerio) were exposed from fertilization until 64 days post-fertilization (dpf) to environmentally relevant (3 and 10ng/L) concentrations of the synthetic estrogen 17α-ethinylestradiol (EE2). The exposure duration covered the period of initial thymus differentiation to maximum growth. Thymus development was assessed by histological and morphometric (thymus area) analysis, thymocyte number, and transcript levels of thymocyte marker genes. Additionally, transcript levels of the estrogen receptors (esr1 and esr2a) were determined. The EE2 exposure altered sexual development (gonad differentiation, transcript levels of hepatic vitellogenin and estrogen receptors) of zebrafish, as expected. At the same time, the EE2 treatment reduced the thymus growth (thymus area, thymocyte number) and transcript levels of thymus marker genes. The expression of the thymic estrogen receptors responded to the EE2 exposure but in a different pattern than the hepatic estrogen receptors. After the 64-day-exposure period, the juvenile fish were transferred into clean water for another 95 days to assess the reversibility of EE2-induced effects. The thymic alterations were found to be reversible in female zebrafish but persisted in males. The present study provides the first evidence that the development of the fish adaptive immune system is sensitive to EEDCs, and that this takes place at concentrations similar to those that disrupt sexual development.

Modeling estrogenic activity in streams throughout the Potomac and Chesapeake Bay watersheds.

Endocrine-disrupting compounds (EDCs), specifically estrogenic endocrine-disrupting compounds, vary in concentration and composition in surface waters under the influence of different landscape sources and landcover gradients. Estrogenic activity in surface waters may lead to adverse effects in aquatic species at both individual and population levels, often observed through the presence of intersex and vitellogenin induction in male fish. In the Chesapeake Bay Watershed, located on the mid-Atlantic coast of the USA, intersex has been observed in several sub-watersheds where previous studies have identified specific landscape sources of EDCs in tandem with observed fish health effects. Previous work in the Potomac River Watershed (PRW), the largest basin within the Chesapeake Bay Watershed, was leveraged to build random forest regression models to predict estrogenic activity at unsampled reaches in both the Potomac River and larger Chesapeake Bay Watersheds (CBW). Model outputs including important variables, partial dependence plots, and predicted values of estrogenic activity at unsampled reaches provide insight into drivers of estrogenic activity at different seasons and scales. Using the US Environmental Protection Agency effects-based threshold of 1.0ng/L 17 β-estradiol equivalents, catchments predicted to exceed this value were categorized as at risk for adverse effects from exposure to estrogenic compounds and evaluated relative to healthy watersheds and recreation access locations throughout the PRW. Results show immediate catchment scale models are more reliable than upstream models, and the best predictive variables differ by season and scale. A small percentage of healthy watersheds (< 13%) and public access sites were classified as at risk using the "Total" (annual) model in the CBW. This study is the first Potomac River Watershed assessment of estrogenic activity, providing a new foundation for future risk assessment and management design efforts, with additional context provided for the entire Chesapeake Bay Watershed.

Analytical determination of oestrogenic endocrine disruptors: the method of choice for wastewater treatment plant effluents

Environmental context Endocrine disrupting compounds (EDCs) are among the most recently targeted micropollutants detected in wastewater treatment plant (WWTP) effluents and in aquatic environments. There is a need for the development of robust analytical methods for most relevant estrogenic EDCs. This study provides optimisation of analytical techniques and addresses several relevant aspects that are often overlooked in the literature. The method was finally successfully employed for the analysis of WWTP effluents. Abstract Two analytical approaches – liquid chromatography–tandem mass spectrometry (LC-MS/MS) and gas chromatography–tandem mass spectrometry (GC-MS/MS) methods – were compared for the simultaneous determination of the 19 most important oestrogenic endocrine disrupting chemicals (EDCs), such as 17β-oestradiol, oestrone, 17α-ethinyloestradiol, bisphenol A and triclosan in wastewater treatment plant effluents. To lower the instrument limits of detection (ILODs), a derivatisation step preceded detection in both methods. The stability, sensitivity and ease of use of dansylation (Dns) for LC-MS/MS and trimethylsilylation (TMS) for GC-MS/MS derivatives were evaluated before method validation. TMS derivatisation products were highly unstable over time. Parameters such as susceptibility to matrix effects and the stability of monodansylated and didansylated derivatisation products of phytohormones are discussed. Lower ILODs of highly potent EDCs (0.11 ng mL−1 for 17β-oestradiol, 0.01 ng mL−1 for 17α-ethinyloestradiol and 0.22 ng mL−1 for oestrone) and stability of derivatisation products within 7 days were achieved using LC–MS/MS; therefore, further validation of this method at environmentally relevant concentrations was conducted. The method limits of detection (MLODs) met the requirements of the European Union defined in Directive 2008/105/ES for 17α-ethinyloestradiol (0.035 ng L−1) and 17β-oestradiol (0.4 ng L−1). Twenty samples of wastewater treatment plant effluent from the Czech Republic were screened using LC-MS/MS. Fifteen of the EDCs were detected in at least one sample. The most abundant EDCs were bisphenol A, with a concentration up to 1107 ng L−1, and triclosan, with a concentration up to 76 ng L−1. No seasonal trend between late spring and autumn samples was observed in the frequency or quantity of analytes.

Ecological risk assessment of environmental stress and bioactive chemicals to riverine fish populations: An individual-based model of smallmouth bass Micropterus dolomieu✰

• A modeling framework is developed to assess population-level effects of temperature, flow and chemicals. • The framework includes three modules: biological model module, effect module and population model. • The framework is demonstrated with an individual-based model of smallmouth bass. • Warm summer water temperatures and year-round high flows have the most severe impacts. • Exposure to estrogenic endocrine disrupting compounds substantially reduces population size. Ecological risk assessments play an important role in environmental management and decision-making. Although empirical measurements of the effects of habitat changes and chemical exposure are often made at molecular and individual levels, environmental decision-making often requires the quantification of management-relevant, population-level outcomes. In this study, we generalized a modeling framework to evaluate population-level ecological risk of environmental stress and bioactive chemicals. The modeling framework includes (1) a biological model module that incorporates complex and interacting biological and ecological processes, and environmental stochasticity, (2) an effect module that links the impacts of environmental changes and chemical exposure to individual characteristics, and (3) a population module that makes decisions on the choice of population-level properties to best capture the effects and thus to track in the model based on the target species and the research and management interest. This framework is a 3-module procedure that provides an alternative way for researchers to organize, present and communicate the risk assessment modeling studies. To demonstrate this framework, we used a socioeconomically important riverine fish species, smallmouth bass Micropterus dolomieu , as the model species. We developed an individual-based model as the biological model module. We evaluated the impacts of changing water temperature and flow regimes, and the impacts of exposure to estrogenic endocrine disrupting compounds (EEDC) on smallmouth bass populations in the Chesapeake Bay Watershed, USA. Warm summer water temperatures and year-round high flows had the most severe impacts on the smallmouth bass population. An increase in exposure level to EEDC, both year-round and in summer months, substantially reduced population size, spawner and recruit abundance, and the proportion of quality-length individuals. Acute exposure to EEDC was more detrimental to the population than chronic exposure. Acute exposure during spawning season had the most severe impacts. This modeling framework can be extended to other species, environmental factors and chemicals, and can be used to inform management and conservation decisions.

17α-ethinylestradiol and 4-tert-octylphenol concurrently disrupt the immune response of common carp

The aquatic environment is massively polluted with endocrine-disrupting compounds (EDCs) including synthetic estrogens (e.g. 17α-ethinylestradiol, EE2) and alkylphenols (e.g. 4-tert-octylphenol, 4t-OP). A major mechanism of action for estrogenic EDCs is their interaction with estrogen receptors and consequently their modulation of the action of enzymes involved in steroid conversion e.g. aromatase CYP19.We now studied the effects of EE2 and 4t-OP on the anti-bacterial immune response of common carp. We investigated effects on the number/composition of inflammatory leukocytes and on the gene expression of mediators that regulate inflammation and EDC binding. In vitro we found that high concentrations of both EE2 and 4t-OP down-regulated IFN-γ2 and IFN-γ-dependent immune responses in LPS-stimulated monocytes/macrophages. Similarly, during bacterial infection in fish, in vivo treated with EE2 and 4t-OP, decreased gene expression of il-12p35 and of ifn-γ2 was found in the focus of inflammation. Moreover, during A. salmonicida-induced infection in EE2-treated carp, but not in fish fed with 4t-OP-treated food, we found an enhanced inflammatory reaction manifested by high number of inflammatory peritoneal leukocytes, including phagocytes and higher expression of pro-inflammatory mediators (inos, il-1β, cxcl8_l2). Furthermore, in the liver, EE2 down-regulated the expression of acute phase proteins: CRPs and C3. Importantly, both in vitro and in vivo, EDCs altered the expression of estrogen receptors: nuclear (erα and erβ) and membrane (gpr30). EDCs also induced up-regulation of the cyp19b gene.Our findings reveal that contamination of the aquatic milieu with estrogenic EDCs, may considerably violate the subtle and particular allostatic interactions between the immune response and endogenous estrogens and this may have negative consequences for fish health.

Long-term exposure to low 17α-ethinylestradiol (EE2) concentrations disrupts both the reproductive and the immune system of juvenile rainbow trout, Oncorhynchus mykiss

Estrogenic endocrine disrupting compounds (EEDCs), such as ethinylestradiol (EE2), are well studied for their impact on the reproductive system of fish. EEDCs may also impact the immune system and, as a consequence, the disease susceptibility of fish. It is currently not yet known whether the low concentrations of EEDCs that are able to disrupt the reproductive system of trout are effective in disrupting the immune system and the fish host resistance towards pathogens, too, or whether such immunodisruptive effects would occur only at higher EEDC concentrations. Therefore, in the present study we compare the effect thresholds of low 17α-ethinylestradiol concentrations (1.5 and 5.5 EE2 ng/L) on the reproductive system, the immune system, the energy expenditures and the resistance of juvenile rainbow trout (Oncorhynchus mykiss) against the parasite Tetracapsuloides bryosalmonae – the etiological agent of proliferative kidney disease (PKD) of salmonids. The parasite infection was conducted without injection and under low pathogen exposure concentrations. The disease development was followed over 130 days post infection – in the presence or absence of EE2 exposure. The results show that the long-term EE2 exposure affected, at both concentrations, reproductive parameters like the mRNA levels of hepatic vitellogenin and estrogen receptors. At the same concentrations, EE2 exposure modulated the immune parameters: mRNA levels of several immune genes were altered and the parasite intensity as well as the disease severity (histopathology) were significantly reduced in EE2-exposed fish compared to infected control fish. The combination of EE2 exposure and parasite infection was energetically costly, as indicated by the decreased values of the swim tunnel respirometry. Although further substantiation is needed, our findings suggest that EE2 exerts endocrine disruptive and immunomodulating activities at comparable effect thresholds, since reproductive and immune parameters were affected by the same, low EE2 concentrations.

Hypothetical roadmap towards endometriosis: prenatal endocrine-disrupting chemical pollutant exposure, anogenital distance, gut-genital microbiota and subclinical infections.

Endometriosis is a gynaecological hormone-dependent disorder that is defined by histological lesions generated by the growth of endometrial-like tissue out of the uterus cavity, most commonly engrafted within the peritoneal cavity, although these lesions can also be located in distant organs. Endometriosis affects ~10% of women of reproductive age, frequently producing severe and, sometimes, incapacitating symptoms, including chronic pelvic pain, dysmenorrhea and dyspareunia, among others. Furthermore, endometriosis causes infertility in ~30% of affected women. Despite intense research on the mechanisms involved in the initial development and later progression of endometriosis, many questions remain unanswered and its aetiology remains unknown. Recent studies have demonstrated the critical role played by the relationship between the microbiome and mucosal immunology in preventing sexually transmitted diseases (HIV), infertility and several gynaecologic diseases. In this review, we sought to respond to the main research question related to the aetiology of endometriosis. We provide a model pointing out several risk factors that could explain the development of endometriosis. The hypothesis arises from bringing together current findings from large distinct areas, linking high prenatal exposure to environmental endocrine-disrupting chemicals with a short anogenital distance, female genital tract contamination with the faecal microbiota and the active role of genital subclinical microbial infections in the development and clinical progression of endometriosis. We performed a search of the scientific literature published until 2019 in the PubMed database. The search strategy included the following keywords in various combinations: endometriosis, anogenital distance, chemical pollutants, endocrine-disrupting chemicals, prenatal exposure to endocrine-disrupting chemicals, the microbiome of the female reproductive tract, microbiota and genital tract, bacterial vaginosis, endometritis, oestrogens and microbiota and microbiota-immune system interactions. On searching the corresponding bibliography, we found frequent associations between environmental endocrine-disrupting chemicals and endometriosis risk. Likewise, recent evidence and hypotheses have suggested the active role of genital subclinical microbial infections in the development and clinical progression of endometriosis. Hence, we can envisage a direct relationship between higher prenatal exposure to oestrogens or estrogenic endocrine-disrupting compounds (phthalates, bisphenols, organochlorine pesticides and others) and a shorter anogenital distance, which could favour frequent postnatal episodes of faecal microbiota contamination of the vulva and vagina, producing cervicovaginal microbiota dysbiosis. This relationship would disrupt local antimicrobial defences, subverting the homeostasis state and inducing a subclinical inflammatory response that could evolve into a sustained immune dysregulation, closing the vicious cycle responsible for the development of endometriosis. Determining the aetiology of endometriosis is a challenging issue. Posing a new hypothesis on this subject provides the initial tool necessary to design future experimental, clinical and epidemiological research that could allow for a better understanding of the origin of this disease. Furthermore, advances in the understanding of its aetiology would allow the identification of new therapeutics and preventive actions.

Direct and indirect parental exposure to endocrine disruptors and elevated temperature influences gene expression across generations in a euryhaline model fish.

Aquatic organisms inhabiting polluted waterways face numerous adverse effects, including physiological disruption by endocrine disrupting compounds (EDCs). Little is known about how the temperatures associated with global climate change may influence the response of organisms exposed to EDCs, and the effects that these combined stressors may have on molecular endpoints such as gene expression. We exposed Menidia beryllina (inland silversides) to environmentally relevant concentrations (1 ng/L) of two estrogenic EDCs (bifenthrin and 17α-ethinylestradiol; EE2) at 22 °C and 28 °C. We conducted this experiment over multiple generations to better understand the potential effects to chronically exposed populations in the wild. We exposed adult parental fish (F0) for 14 days prior to spawning of the next generation. F1 larvae were then exposed from fertilization until 21 days post hatch (dph) before being transferred to clean water tanks. F1 larvae were reared to adulthood, then spawned in clean water to test for further effects of parental exposure on offspring (F2 generation). Gene expression was quantified by performing qPCR on F0 and F1 gonads, as well as F1 and F2 larvae. We did not detect any significant differences in the expression of genes measured in the parental or F1 adult gonads. We found that the 28 °C EE2 treatment significantly decreased the expression of nearly all genes measured in the F1 larvae. This pattern was transferred to the F2 generation for expression of the follicle-stimulating hormone receptor (FSHR) gene. Expression of 17β-hydroxysteroid dehydrogenase (17β-HSD) and G protein-coupled receptor 30 (GPR30) revealed changes not measured in the previous generation. Effects of the bifenthrin treatments were not observed until the F2 generation, which were exposed to the chemicals indirectly as germ cells. Our results indicate that effects of EDCs and their interactions with abiotic factors, may not be adequately represented by singular generation testing. These findings will contribute to the determination of the risk of EDC contamination to organisms inhabiting contaminated waterways under changing temperature regimes.

Atrazine affects craniofacial chondrogenesis and axial skeleton mineralization in zebrafish ( Danio rerio).

Atrazine is a commonly used herbicide that has previously been implicated as an endocrine-disrupting compound. Previous studies have shown that estrogenic endocrine-disrupting compounds affect the development of the heart, cartilage, and bone in zebrafish ( Danio rerio). To determine whether atrazine has effects similar to other endocrine disruptors, zebrafish embryos were treated with a range of atrazine concentrations. Atrazine treatment at a low concentration of 0.1 µM resulted in significant differences in craniofacial cartilage elements, while concentrations ≥1 µM led to decreased survival and increased heart rates. Fish treated with ≥1 µM atrazine also developed with delayed vertebrae mineralization. Higher concentrations of atrazine caused gross craniofacial defects and decreased hatching rates. Further studies into the molecular pathways disrupted in these developmental processes could shed light on a link between endocrine-disrupting compounds and developmental abnormalities.

Testicular oocytes in smallmouth bass in northeastern Minnesota in relation to varying levels of human activity.

Testicular oocytes (TOs) have been found in black bass (Micropterus spp.) from many locations in North America. The presence of TOs is often assumed to imply exposure to estrogenic endocrine disrupting compounds (EDCs); however, a definitive causal relationship has yet to be established, and TO prevalence is not consistently low in fish from areas lacking evident EDC sources. This might indicate any of a number of situations: 1) unknown or unidentified EDCs or EDC sources, 2) induction of TOs by other stressors, or 3) testicular oocytes occurring spontaneously during normal development. In the present study, we analyzed TO occurrence in smallmouth bass (Micropterus dolomieu) from 8 populations in northeastern Minnesota watersheds with differing degrees of human development and, hence, presumed likelihood of exposure to anthropogenic chemicals. Three watersheds were categorized as moderately developed, based on the presence of municipal wastewater discharges and higher human population density (4-81 per km2 ), and 5 watersheds were minimally developed, with very low human population density (0-1 per km2 ) and minimal built environment. Testicular tissues from mature fish were evaluated using a semiquantitative method that estimated TO density, normalized by cross-sectional area. Testicular oocyte prevalence and density among populations from moderately developed watersheds was higher than in populations from minimally developed watersheds. However, TO prevalence was unexpectedly high and variable (7-43%) in some populations from minimally developed watersheds, and only weak evidence was found for a relationship between TO density and watershed development, suggesting alternative or more complex explanations for TO presence in smallmouth bass from this region. Environ Toxicol Chem 2017;36:3424-3435. © 2017 SETAC.

Combined effects of increased temperature and endocrine disrupting pollutants on sex determination, survival, and development across generations.

Understanding the combined effects of anthropogenic impacts such as climate change and pollution on aquatic ecosystems is critical. However, little is known about how predicted temperature increases may affect the activity of endocrine disrupting compounds (EDCs), particularly in species with plasticity in sex determination. We investigated the effects of a concomitant increase in temperature and exposure to estrogenic EDCs on reproduction and development in an estuarine model organism (Menidia beryllina) across multiple generations. Parents (P) were exposed to environmental levels of the estrogenic insecticide bifenthrin or ethinylestradiol (EE2) at 22 °C and 28 °C for 14 days prior to the initiation of spawning trials. Embryos in the F1 generation were exposed to EDCs until 21 days post hatch (dph), reared to adulthood in clean water at elevated temperatures, and spawned. F1 sex ratios were significantly influenced by elevated temperature and EDCs, potentially altering adaptive development. We also observed fewer viable offspring and increased developmental deformities in the F1 and F2 generations, with a greater impact on F2 juveniles. These findings enhance our understanding of responses to EDCs in the context of climate change and may demonstrate heritable effects. Our study represents the first multigenerational assessment of elevated temperatures in combination with environmentally relevant concentrations of commonly detected endocrine disruptors in a model vertebrate species.

Removal of Six Estrogenic Endocrine-Disrupting Compounds (EDCs) from Municipal Wastewater Using Aluminum Electrocoagulation

Conventional wastewater treatment plant (WWTP) processes are primarily designed to reduce the amount of organic matter, pathogens, and nutrients from the incoming influent. However, these processes are not as effective in reducing the concentrations of micropollutants, including endocrine-disrupting compounds (EDCs), which notoriously evade traditional wastewater treatment technologies and are found even in tertiary-treated effluent. For WWTPs practicing deep-well injection or surface-water discharge, EDCs in the treated effluent are discharged into groundwater or the aquatic environment where humans and wildlife may potentially suffer the effects of chemical exposure. In the current laboratory-scale study, we tested a bench-top electrocoagulation (EC) unit utilizing aluminum blades for the removal of six estrogenic EDCs [estrone (E1), 17β-estradiol (E2), estriol (E3), 17α-ethinylestradiol (EE2), bisphenol-A (BPA), and nonylphenol (NP)]. Samples of municipal wastewater influent and tertiary-treated effluent were spiked with the six EDCs in order to test the removal efficiency of the EC unit. The mean concentration of each EDC component was statistically lower after EC treatment (removal range = 42%–98%). To our knowledge, this is the first study to investigate aluminum electrocoagulation for removal of these specific EDCs, including nonylphenol (without the ethoxylate chain), as well as natural and synthetic estrogens.

Behaviour of estrogenic endocrine-disrupting chemicals in permeable carbonate sands.

The remediation of four estrogenic endocrine-disrupting compounds (EDCs), estrone (E1), estradiol (E2), ethinylestradiol (EE2) and estriol (E3), was measured in saturated and unsaturated carbonate sand-filled columns dosed with wastewater from a sewage treatment plant. The estrogen equivalency (EEQ) of inlet wastewater was 1.2 ng L(-1) and was remediated to an EEQ of 0.5 ng L(-1) through the unsaturated carbonate sand-filled columns. The high surface area of carbonate sand and associated high microbial activity may have assisted the degradation of these estrogens. The fully saturated sand columns showed an increase in total estrogenic potency with an EEQ of 2.4 ng L(-1), which was double that of the inlet wastewater. There was a significant difference (P < 0.05) in total estrogenic potency between aerobic and anaerobic columns. The breakdown of conjugated estrogens to estrogenic EDCs formed under long residence time and reducing conditions may have been responsible for the increase in the fully saturated columns. This may also be explained by the desorption of previously sorbed estrogenic EDCs. The effect of additional filter materials, such as basalt sediment and coconut fibre, on estrogenic EDC reduction was also tested. None of these amendments provided improvements in estrogen remediation relative to the unamended unsaturated carbonate sand columns. Aerobic carbonate sand filters have good potential to be used as on-site wastewater treatment systems for the reduction of estrogenic EDCs. However, the use of fully saturated sand filters, which are used to promote denitrification, and the loss of nitrogen as N2 were shown to cause an increase in EEQ. The potential for the accumulation of estrogenic EDCs under anaerobic conditions needs to be considered when designing on-site sand filtration systems required to reduce nitrogen. Furthermore, the accumulation of estrogens under anaerobic conditions such as under soil absorption systems or leachate fields has the potential to contaminate groundwater especially when the water table levels fluctuate.